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Next-generation Sequencing

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....

RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...

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Related Experiment Video

Updated: Jun 10, 2025

Multiplexed Analysis of Retinal Gene Expression and Chromatin Accessibility Using scRNA-Seq and scATAC-Seq

Multiplexed Analysis of Retinal Gene Expression and Chromatin Accessibility Using scRNA-Seq and scATAC-Seq

Published on: March 12, 2021

NucBalancer: streamlining barcode sequence selection for optimal sample pooling for sequencing.

Saurabh Gupta^1,2, Ankur Sharma^1,2,3

¹Harry Perkins Institute of Medical Research, QEII Medical Centre and Centre for Medical Research, 6 Verdun Street, Nedlands, Perth, Western Australia, 6009, Australia.

Gigabyte (Hong Kong, China)

|October 21, 2024

Summary

This summary is machine-generated.

NucBalancer is a new R-based tool that optimizes barcode selection for next-generation sequencing (NGS). It ensures accurate sample pooling and demultiplexing across diverse platforms, enhancing data reliability.

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Area of Science:

Genomics
Bioinformatics

Background:

Next-generation sequencing (NGS) technologies are rapidly evolving, necessitating adaptable and cost-effective tools.
New sequencing platforms present challenges in sample preparation, pooling, and demultiplexing due to unique requirements.
Library conversion between platforms requires efficient barcode balancing to maintain data integrity.

Purpose of the Study:

To introduce NucBalancer, a Shiny app designed for optimal barcode sequence selection in NGS.
To address nucleotide composition challenges for specific sequencers (G400, T7) and broader NGS applications.
To facilitate adaptation of libraries, such as 10x Genomics, for various sequencing platforms.

Main Methods:

Development of NucBalancer, a versatile Shiny application implemented in R.
Algorithm designed for optimal selection of barcode sequences.
Functionality to balance nucleotide composition and sample concentrations for pooling.

Main Results:

NucBalancer efficiently balances nucleotide composition and sample concentrations, reducing biases.
The tool enhances the reliability of NGS data across different sequencing platforms.
Enables adaptation of libraries for diverse sequencing needs, including single-cell genomics.

Conclusions:

NucBalancer provides a crucial solution for barcode balancing in the dynamic NGS landscape.
Its adaptability supports various sequencing technologies and applications, improving data accuracy.
The tool is invaluable for ensuring effective sample pooling and demultiplexing on any platform.